Project Summary Transfusions of donated blood are the safest and most effective way to treat a patient with severe blood loss. However, there are many situations (e.g. military battlefields, remote areas, developing countries) in which blood is unavailable because it must be constantly refrigerated. The purpose of this project is to develop a ?blood substitute? that does not require refrigeration and can be deployed to save lives in remote areas. Most previous blood substitutes have been based on purified human hemoglobin, since it is a protein that does not require refrigeration and it is responsible for the most vital function of human blood ? oxygen transport. Unfortunately, clinical trials have revealed that blood substitutes based on human hemoglobin cause many different serious side effects, including heart attack and stroke. The goal of this project is to find an alternative hemoglobin that can effectively transport oxygen without causing the side effects observed with human hemoglobin. Specifically, the efficacy and safety of 2 invertebrate hemoglobins with unique properties will be investigated by transfusing them into mice. The effects of long term storage (>1-2 weeks) at high temperatures (70-140oF) on each of the hemoglobins will also be investigated to determine if they can be deployed to remote areas where donated blood is unavailable. Finally, the hemoglobins will be chemically modified to increase their stability and a freeze drying protocol will be developed to produce a blood substitute that is ultra- stable and ultra-portable. This work will enhance the research environment at Villanova University by establishing a valuable collaboration with experienced researchers at the Children's Hospital of Philadelphia. This project will also provide support for at least 1 graduate and 12 undergraduate students to gain valuable research experience in the field of drug discovery and development by working at both Villanova and CHOP.